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Hibiscus rosasinensis, medicinal plant known for its various medicinal properties. The present study was to identify and characterize the flavonoid compound from the petals of Hibiscus rosa sinensis. The antioxidant compounds were isolated by Silica Gel G Column chromatography. From the fractions, C5 shows the highest antioxidant activity compared to C3 & C4 compounds. DNA damage protection activity results indicated that two major bands super coiled DNA and open circular DNA was protected by the presence of different concentration (5 µg, 10 µg, 15 µg & 20 µg) of C5 compound isolated from the petals of Hibiscus rosa sinensis. IR absorptions peaks reveals the presence of functional groups alcohols, phenols, α,β unsaturated aldehyes, ketones and alkanes. The structure of compound was characterized as Hibiscetin-3-glucoside (C21H20O14) by Mass spectroscopy and NMR. The study emphasized flavanoid compounds have effective scavenging activities which may be due to its phenolics and flavonoid contents and can be used as anticancer agents. Keywords: Flavanoid, Antioxidant, DNA protector activity, IR, NMR, Hibiscetin-3-glucoside